KR102659269B1 - Tetragonia tetragonioides extract showing whitening and anti-inflammatory activity - Google Patents

Abstract

The present invention provides an extract of Prickly pear sinensis characterized by skin whitening activity, antioxidant activity, and anti-inflammatory activity.

Description

{Tetragonia tetragonioides extract showing whitening and anti-inflammatory activity}

The present invention relates to an extract of Prickly pear sinensis that exhibits whitening and anti-inflammatory activities.

A person's skin color is affected by the presence or absence of pigments such as skin melanin, hemoglobin, or carotenoids produced by melanocytes, which are pigment cells present in the epidermis, and the thickness and reflectivity of the skin. Among these, melanin pigment, which is the most important factor in determining skin color, is made by converting tyrosin, a type of amino acid that normally exists in the human body, into DOPA by tyrosinase, an enzyme present in melanocytes, and a series of Through a complex oxidation process, melanin, a dark brown polymer, is ultimately produced.

Melanin synthesis is promoted by factors such as exposure to ultraviolet rays, melanoma, and hyperpigmentation disease. Recently, a molecular biological approach has been attempted as it has been reported that the expression and synthesis of tyrosinase, TRP-1, and TRP-2 are directly involved in melanin synthesis.

On the other hand, the inflammatory response occurs when tissue (cells) are damaged or infected with external infectious agents (bacteria, mold, viruses, various types of allergens), various inflammatory mediators and immune cells in local blood vessels and body fluids are involved, resulting in enzyme activation, It shows a series of complex physiological reactions such as secretion of inflammatory mediators, fluid infiltration, cell migration, and tissue destruction, as well as external symptoms such as erythema, edema, fever, and pain. In normal cases, the inflammatory response removes external infectious agents and regenerates damaged tissues to restore the function of the organism. However, if the inflammatory response occurs excessively or continuously due to antigens not being removed or internal substances as the cause, it actually promotes mucosal damage, resulting in In some cases, it leads to diseases such as cancer. Recently, research is being actively conducted to use natural substances rather than artificial substances in various fields such as substances, functional foods, and functional cosmetics to prevent and treat various diseases such as inflammatory diseases as mentioned above.

Meanwhile, Tetragonia tetragonioides is a perennial herb belonging to the Aizoaceae family, also known as sea lettuce or New Zealand spinach. It is 40-60 cm tall, has fleshy protuberances, has many branches at the bottom, and the stem stands at an angle or extends to the side. It falls to the ground and then gradually stands up, growing to a height of about 50 cm. It branches out slightly, attaches itself to the ground in a circle from one root, and spreads out in all directions. It is also distributed in New Zealand, China, Japan, South Asia, Australia, and South America, and has strong vitality, so it grows well even in extremely barren and moisture-deprived places such as gravel fields or rock crevices. It is an edible crop that is used as a raw material in salads and can also be consumed as a vegetable when boiled. Bunhaengcho is said to be a herb in traditional Chinese medicine, and the herb has been used to treat enteritis, sepsis, carbuncle, and wind fever due to its effectiveness in clearing heat, detoxifying, blowing wind, and soybean. To date, β-carctene, diterpene, flavone, cerebrosides, oxilic acid, sterylglucoside, and polysaccharide have been reported as components isolated from this plant, and anti-inflammatory and anti-ulcer activities have been reported. However, to date, no research has been conducted on the skin physiological activity of Bunhaengcho, so more research on the skin physiological activity of Bunhaengcho is needed.

The present invention has been devised to solve the above problems, and the purpose of the present invention is to provide an extract of Prickly pear sinensis that exhibits whitening activity and anti-inflammatory activity.

In addition, another object of the present invention is to provide a cosmetic composition for whitening containing an extract of fennel root as an active ingredient.

The object of the present invention is not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to achieve the above object, the present invention

Provided is an extract of Prickly pear sinensis, characterized in that it exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the present invention

Provided is a composition comprising, as an active ingredient, an extract of Prickly pear herbacea, which exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the composition is characterized in that it contains 200-500 μg/ml of 70% (v/v) ethanol extract of the dried product of Bunhaengcho as an active ingredient.

In addition, the extract of Bunhaengcho,

Drying the raw material of Tetragonia tetragonioides at a temperature of 40-60°C;

Preparing dried herbaceous herb powder by grinding dried herbaceous root with a grinder;

Immersing the powder in ethanol and leaching it by stirring at a temperature of 20-30°C, filtering the leached material, and repeating the leaching and filtration 2-3 times; and

It is characterized in that it is manufactured by performing the steps of concentrating the filtrate obtained by filtration under reduced pressure and freeze-drying.

In addition, the present invention

Provided is a cosmetic composition for skin whitening that contains an extract of Bunchesia chinensis as an active ingredient and exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the present invention

Provided is a health functional food composition for the prevention or improvement of melanin hyperpigmentation disease, which contains an extract of fennel root as an active ingredient and exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the present invention

Provided is a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation disease, which contains an extract of Bunchesia chinensis as an active ingredient and exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the present invention

1-5 parts by weight of the above-mentioned Bunhaengcho extract, 0.5-1.5 parts by weight of a fruit extract mixed with tangerine extract, kiwi extract, pear extract, and orange extract in a weight ratio of 3:3:2:2, honeysuckle extract, and honeysuckle extract. and 0.5-1.5 parts by weight of flower extract mixed with peony flower extract at a weight ratio of 1:1:1, 0.5-1.5 parts by weight of grapefruit-derived exosomes, fermented corn extract, and fermented centella asiatica extract mixed at a weight ratio of 1:1. 0.5-1.5 parts by weight of fermented extract, 1-3 parts by weight of niacinamide, 0.5-1.5 parts by weight of arginine, 0.5-1.5 parts by weight of phenylalanine, 0.5-1.5 parts by weight of 1,2-hexanediol, 0.5 parts by weight of pentylene glycol. -1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1-3 parts by weight, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05- Provided is a cosmetic composition comprising 0.2 parts by weight, polyglyceryl-6 stearate 0.05-0.2 parts by weight, glutamic acid 0.3-0.7 parts by weight, and carrot water 68-72 parts by weight.

In addition, the present invention

Drying the raw material of Tetragonia tetragonioides at a temperature of 40-60°C; Preparing dried herbaceous herb powder by grinding dried herbaceous root with a grinder; Immersing the powder in ethanol and leaching it by stirring at a temperature of 20-30°C, filtering the leached material, and repeating the leaching and filtration 2-3 times; and concentrating the filtrate obtained by filtration under reduced pressure and freeze-drying; preparing an extract of Bunhaengcho, including;

Washing, freeze-drying, and pulverizing tangerines, kiwis, pears, and oranges to prepare tangerine powder, kiwi powder, pear powder, and orange powder; The prepared fruit powder mixed with tangerine powder, kiwi powder, pear powder, and orange powder at a weight ratio of 3:3:2:2 was mixed with a mixed solvent of ethanol and hexane at a volume ratio of 1:1 and a weight ratio of 1:9. mixing and extracting at a temperature of 28-32°C for 22-26 hours to prepare an extract; And filtering the extract through filter paper, concentrating and freeze-drying the filtered filtrate under reduced pressure at a temperature of -48-52°C; Preparing a fruit extract comprising a;

Preparing honeysuckle flower powder, Yangha flower powder, and peony flower powder by washing, freeze-drying, and pulverizing honeysuckle flowers, Yangha flowers, and peony flowers; 20 g of the prepared mixed powder of honeysuckle flower powder, Yangha flower powder, and peony flower powder mixed at a weight ratio of 1:1:1 was mixed with 200 ml of water and stirred for 6-10 minutes at a temperature of 78-82°C. performing primary hydrothermal extraction for a period of time; After the first hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a first mixed extract; After the first hot water extraction, filter with filter paper, mix the remaining solids with water again so that the mixing ratio of solids and water is 0.1 g/ml, and do the second extraction at a temperature of 118-122°C for 10-14 hours while stirring. performing hot water extraction; After secondary hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a secondary mixed extract; After secondary hot water extraction and filtering with filter paper, the remaining solids are mixed with water again so that the mixing ratio of solids and water is 0.1 g/ml, and then mixed with [BMIm][TFSI](1-Butyl-3-methylimidazoliumbis). Adding 1-2 g of trifluoromethylsulfonyl)imide) and performing a third hot water extraction at a temperature of 98-102°C for 6-10 hours while stirring; After the third hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a third mixed extract; And mixing the first mixed extract, second mixed extract, and third mixed extract; Preparing a flower extract comprising a;

Washing the grapefruit and pretreating it at a temperature of 18-22°C and a pressure of 140-160 MPa for 3-7 minutes; Preparing grapefruit juice by squeezing pretreated grapefruit; Centrifuging the grapefruit juice to obtain a supernatant containing exosomes; Preparing a lyophilisate by lyophilizing the supernatant containing the exosomes; Forming an aqueous two-phase system using polyethylene glycol/dextran in the freeze-dried product; And obtaining a lower layer liquid in which exosomes are concentrated in the aqueous two-phase system; Preparing grapefruit-derived exosomes comprising;

Preparing corn powder and Centella asiatica powder by washing corn grains and Centella asiatica with boiled water and pulverizing them; A mixture was prepared by mixing 23-27 parts by weight of corn powder, 23-27 parts by weight of Centella asiatica powder, and 58-62 parts by weight of water, and 1 part by weight of Saccharomyces cerevisiae as a yeast was added to the mixture. Fermentation at a temperature of 38-42°C for 34-38 hours; Preparing an extract by mixing the fermented mixture and butylene glycol and stirring for 4-6 days at a temperature of 28-32°C; and filtering and concentrating the extract under reduced pressure and then freeze-drying the extract. Preparing a fermentation extract including; and

1-5 parts by weight of the Bunhaengcho extract, 0.5-1.5 parts by weight of the fruit extract, 0.5-1.5 parts by weight of the flower extract, 0.5-1.5 parts by weight of the grapefruit-derived exosomes, 0.5-1.5 parts by weight of the fermented extract, niacinamide 1 -3 parts by weight, arginine 0.5-1.5 parts by weight, phenylalanine 0.5-1.5 parts by weight, 1,2-hexanediol 0.5-1.5 parts by weight, pentylene glycol 0.5-1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight Parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1-3 parts by weight, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05-0.2 parts by weight, polyglyceryl-6 stearate 0.05 It provides a method for producing a cosmetic composition comprising - mixing 0.2 parts by weight, 0.3-0.7 parts by weight of glutamic acid, and 68-72 parts by weight of carrot water.

According to the present invention, an extract of Prickly pear sinensis that exhibits whitening activity, antioxidant activity, and anti-inflammatory activity is provided. In addition, according to the present invention, a whitening composition is provided containing an extract of Peach blossom root as an active ingredient.

Figure 1 is a photograph showing the manufacturing process of an extract according to an embodiment of the present invention.
Figure 2 is a graph showing the total polyphenol content of the extract according to an embodiment of the present invention,
Figure 3 is a graph showing the ABTS radical scavenging activity of the extract according to an embodiment of the present invention;
Figure 4 is a graph showing the DPPH radical scavenging activity of the extract according to an embodiment of the present invention;
Figure 5 is a graph showing the effect of the extract on cell viability in RAW 264.7 cells induced with LPS according to an embodiment of the present invention;
Figure 6 is a graphical representation of the effect of the extract on NO production in RAW 264.7 cells induced with LPS according to an embodiment of the present invention;
Figure 7 graphically shows the effect of the hexapeptide of the extract according to an example of the present invention on the melanin content in α-MSH stimulated B16F10 cells.

Hereinafter, various embodiments will be described in more detail. The embodiments described herein may be modified in various ways. Specific embodiments may be described in detail in the detailed description. However, the specific embodiments disclosed are only intended to facilitate understanding of the various embodiments. Accordingly, the technical idea is not limited to the specific embodiments disclosed, and should be understood to include all equivalents or substitutes included in the spirit and technical scope of the invention.

Terms containing ordinal numbers, such as primary, secondary, first, second, etc., may be used to describe various components, but these components are not limited by the above-mentioned terms. The above-mentioned terms are used only for the purpose of distinguishing one component from another.

In this specification, terms such as 'include' or 'have' are intended to designate the presence of features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, but are not intended to indicate the presence of one or more other features. It should be understood that this does not exclude in advance the possibility of the existence or addition of elements, numbers, steps, operations, components, parts, or combinations thereof. When a component is said to be 'connected' or 'connected' to another component, it is understood that it may be directly connected or connected to the other component, but that other components may exist in between. It should be. On the other hand, when a component is mentioned as being 'directly connected' or 'directly connected' to another component, it should be understood that there are no other components in between.

In addition, when describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description thereof is abbreviated or omitted.

This invention

Provided is an extract of Prickly pear sinensis, characterized in that it exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

As a result of researching and testing various physiological activities using the above-described extracts of Bunchesia sinensis, the inventors of the present invention were able to find that the extracts of Bunchesa sinensis of the present invention had excellent whitening activity and that they had antioxidant and anti-inflammatory activities. It has been done.

In addition, the present invention

Provided is a composition comprising, as an active ingredient, an extract of Prickly pear herbacea, which exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

The composition preferably contains 200-500 ㎍/ml of the 70% (v/v) ethanol extract of the dried product of Bunhaengcho as an active ingredient, and more preferably contains 250-450 ㎍/ml as an active ingredient.

The extract of Bunhaengcho,

Drying the raw material of Tetragonia tetragonioides at a temperature of 40-60°C;

Preparing dried herbaceous herb powder by grinding dried herbaceous root with a grinder;

Immersing the powder in ethanol and leaching it by stirring at a temperature of 20-30°C, filtering the leached material, and repeating the leaching and filtration 2-3 times; and

Use the product prepared by concentrating the filtrate obtained by filtration under reduced pressure and freeze-drying it.

By preparing the dried extract and then leaching it instead of leaching it as is, it not only exhibits whitening activity, but also exhibits excellent antioxidant and anti-inflammatory activity.

In addition, the present invention

Provided is a cosmetic composition for skin whitening that contains an extract of Bunchesia chinensis as an active ingredient and exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the present invention

1-5 parts by weight of the above-mentioned Bunhaengcho extract, 0.5-1.5 parts by weight of a fruit extract mixed with tangerine extract, kiwi extract, pear extract, and orange extract in a weight ratio of 3:3:2:2, honeysuckle extract, and honeysuckle extract. and 0.5-1.5 parts by weight of flower extract mixed with peony flower extract at a weight ratio of 1:1:1, 0.5-1.5 parts by weight of grapefruit-derived exosomes, fermented corn extract, and fermented centella asiatica extract mixed at a weight ratio of 1:1. 0.5-1.5 parts by weight of fermented extract, 1-3 parts by weight of niacinamide, 0.5-1.5 parts by weight of arginine, 0.5-1.5 parts by weight of phenylalanine, 0.5-1.5 parts by weight of 1,2-hexanediol, 0.5 parts by weight of pentylene glycol. -1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1-3 parts by weight, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05- Provided is a cosmetic composition comprising 0.2 parts by weight, polyglyceryl-6 stearate 0.05-0.2 parts by weight, glutamic acid 0.3-0.7 parts by weight, and carrot water 68-72 parts by weight.

Tangerines contain yellow beta-cryptoxanthin, which has a virus-fighting effect 5 times higher than beta-carotene, so it has excellent anti-cancer effects, and thanks to the action of vitamin C, it has excellent skin aging prevention and whitening effects. In addition, it is rich in calcium and is known to be effective in dieting by excreting sodium from the body.

Kiwi (Actinidia chinensis) is the fruit of a deciduous vine of the dicotyledonous plant Actinidia family Actinidia and is called kiwi in English because it looks like a kiwi bird. In particular, it is rich in vitamin C, so one fruit is sufficient for the vitamin intake required by one adult per day. It is also rich in folic acid, potassium, vitamin A, calcium, vitamin B6, copper, vitamin E, fiber, and potassium.

Pear (Pyrus pyrifolia) has a moisture content of 85% to 88% by weight, has 51 calories, and its main ingredient is carbohydrate. It is a strongly alkaline food containing 10 to 15% by weight of sugar, and 75% by weight of trace elements such as sodium, potassium, and magnesium. Therefore, pears are known to be very effective in maintaining health by keeping blood neutral.

Orange (Citrus sinensis) is a plant belonging to the tangerine genus and contains ingredients such as vitamin C, vitamin B1, B2, flavonoids, beta cartonin, citric acid, and pectin. It also contains carotene, which is not found in lemons, and other useful ingredients. It contains a lot.

In the present invention, the fruit extract extracted from the tangerine, kiwi, pear, and orange is intended to be applied as a cosmetic composition for skin whitening, and the fruit extract exhibits an excellent skin whitening effect.

Lonicera Caprifolium (Honeysuckle), also called honeysuckle or honeysuckle, is mainly distributed in East Asia. It has antipyretic, detoxifying, antibacterial, and anti-inflammatory effects and has been used as a treatment for inflammation and infectious diseases.

Zingiber mioga (Zingiber mioga) is a perennial edible plant belonging to the ginger family and is mainly cultivated in East Asia. In Korea, it is mainly produced in the southern coast and Jeju Island area. Yangha's flower buds have a unique spicy flavor called mioganal and are mainly consumed as seasoned or pickled foods. In a previous study, it was reported that Yanghae extract has the ability to reduce blood glucose concentration by inhibiting carbohydrate hydrolyzing enzymes, and was shown to improve memory in a mouse model with dementia induced by scopolamine. There were also research reports that found it helpful. In addition, research results have been reported on the antioxidant effect and allergic asthma improvement effect of Yanghae extract.

Peony (Paeonia lactiflora, Paeonia albiflora) is a perennial plant of the dicotyledonous plant Peony genus and is mainly cultivated in China. Peony flowers are large in size and have good color, so they were mainly used only for gardening, but recently, various attempts have been made to use peony flowers for skin care purposes.

In the present invention, it is intended to apply flower extracts extracted from the above-mentioned honeysuckle flowers, apricot flowers, and peony flowers as a cosmetic composition for skin whitening, and the application of the flower extract exhibits an excellent skin whitening effect.

Grapefruit (Citrus Paradisi, Grapefruit) is the fruit of the grapefruit tree, belonging to the genus Citrus. Grapefruit is a relatively large tree, has leaves with prominent wing-shaped petioles, blooms fragrant white flowers, and produces large round fruits in small bunches like grapes. The flesh is light yellow, rich in juice, and has a sour yet strong sweet taste with a slightly bitter taste.

In the present invention, grapefruit-derived exosomes are used to apply them as a cosmetic composition for skin whitening, and the grapefruit-derived exosomes are applied to exhibit excellent skin whitening effects.

Corn contains ingredients such as protein, lipid, sugar, fiber, minerals, and vitamins, and is effective in preventing dryness and aging of the skin and increasing resistance to skin eczema. It is the main ingredient in gum disease treatment and has a pharmacological effect in improving teeth. It is also known to be very high. The fiber in corn stimulates the intestines and stimulates intestinal movement, and it contains a lot of vitamin B1, so it is reported to be effective in treating loss of appetite, drowsiness, and lethargy.

Centella asiatica is a perennial creeping herb belonging to the Umbelliferae family. It mainly grows in hot and humid places and is found in wetlands such as Africa, India, and Sri Lanka. In Korea, it mainly lives on Jeju Island and the southern islands, and the madecassoside component in Centella asiatica has long been used to treat skin ulcers and wounds.

In the present invention, the fermentation extract extracted after fermentation using the corn and centella asiatica is intended to be applied as a cosmetic composition for skin whitening, and the application of the fermentation extract shows an excellent skin whitening effect.

Carrots are a vegetable with a high water content containing 90% water, and are rich in nutrients that quickly restore rough skin by containing vitamins A, K and minerals. Additionally, the moisture in carrots has a positive effect on moisturizing the skin. The carrot water is a water-soluble raw material that dissolves the abundant moisture and minerals of carrots, and is generally used instead of purified water used in cosmetic compositions.

In addition, the present invention

Drying the raw material of Tetragonia tetragonioides at a temperature of 40-60°C; Preparing dried herbaceous herb powder by grinding dried herbaceous root with a grinder; Immersing the powder in ethanol and leaching it by stirring at a temperature of 20-30°C, filtering the leached material, and repeating the leaching and filtration 2-3 times; and concentrating the filtrate obtained by filtration under reduced pressure and freeze-drying; preparing an extract of Bunhaengcho, including;

Washing, freeze-drying, and pulverizing tangerines, kiwis, pears, and oranges to prepare tangerine powder, kiwi powder, pear powder, and orange powder; The prepared fruit powder mixed with tangerine powder, kiwi powder, pear powder, and orange powder at a weight ratio of 3:3:2:2 was mixed with a mixed solvent of ethanol and hexane at a volume ratio of 1:1 and a weight ratio of 1:9. mixing and extracting at a temperature of 28-32°C for 22-26 hours to prepare an extract; And filtering the extract through filter paper, concentrating and freeze-drying the filtered filtrate under reduced pressure at a temperature of -48-52°C; Preparing a fruit extract comprising a;

Preparing honeysuckle flower powder, Yangha flower powder, and peony flower powder by washing, freeze-drying, and pulverizing honeysuckle flowers, Yangha flowers, and peony flowers; 20 g of the prepared mixed powder of honeysuckle flower powder, Yangha flower powder, and peony flower powder mixed at a weight ratio of 1:1:1 was mixed with 200 ml of water and stirred for 6-10 minutes at a temperature of 78-82°C. performing primary hydrothermal extraction for a period of time; After the first hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a first mixed extract; After the first hot water extraction, filter with filter paper, mix the remaining solids with water again so that the mixing ratio of solids and water is 0.1 g/ml, and do the second extraction at a temperature of 118-122°C for 10-14 hours while stirring. performing hot water extraction; After secondary hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a secondary mixed extract; After secondary hot water extraction and filtering with filter paper, the remaining solids are mixed with water again so that the mixing ratio of solids and water is 0.1 g/ml, and then mixed with [BMIm][TFSI](1-Butyl-3-methylimidazoliumbis). Adding 1-2 g of trifluoromethylsulfonyl)imide) and performing a third hot water extraction at a temperature of 98-102°C for 6-10 hours while stirring; After the third hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a third mixed extract; And mixing the first mixed extract, second mixed extract, and third mixed extract; Preparing a flower extract comprising a;

Washing the grapefruit and pretreating it at a temperature of 18-22°C and a pressure of 140-160 MPa for 3-7 minutes; Preparing grapefruit juice by squeezing pretreated grapefruit; Centrifuging the grapefruit juice to obtain a supernatant containing exosomes; Preparing a lyophilisate by lyophilizing the supernatant containing the exosomes; Forming an aqueous two-phase system using polyethylene glycol/dextran in the freeze-dried product; And obtaining a lower layer liquid in which exosomes are concentrated in the aqueous two-phase system; Preparing grapefruit-derived exosomes comprising;

Preparing corn powder and Centella asiatica powder by washing corn grains and Centella asiatica with boiled water and pulverizing them; A mixture was prepared by mixing 23-27 parts by weight of corn powder, 23-27 parts by weight of Centella asiatica powder, and 58-62 parts by weight of water, and 1 part by weight of Saccharomyces cerevisiae as a yeast was added to the mixture. Fermentation at a temperature of 38-42°C for 34-38 hours; Preparing an extract by mixing the fermented mixture and butylene glycol and stirring for 4-6 days at a temperature of 28-32°C; and filtering and concentrating the extract under reduced pressure and then freeze-drying the extract. Preparing a fermentation extract including; and

1-5 parts by weight of the Bunhaengcho extract, 0.5-1.5 parts by weight of the fruit extract, 0.5-1.5 parts by weight of the flower extract, 0.5-1.5 parts by weight of the grapefruit-derived exosomes, 0.5-1.5 parts by weight of the fermented extract, niacinamide 1 -3 parts by weight, arginine 0.5-1.5 parts by weight, phenylalanine 0.5-1.5 parts by weight, 1,2-hexanediol 0.5-1.5 parts by weight, pentylene glycol 0.5-1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight Parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1-3 parts by weight, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05-0.2 parts by weight, polyglyceryl-6 stearate 0.05 It provides a method for producing a cosmetic composition comprising - mixing 0.2 parts by weight, 0.3-0.7 parts by weight of glutamic acid, and 68-72 parts by weight of carrot water.

First, the method for producing a cosmetic composition according to the present invention includes the step of preparing an extract of Prickly pear herbacea. The step of preparing the extract of Tetragonia tetragonioides includes drying the raw material of Tetragonia tetragonioides at a temperature of 40-60°C; Preparing dried herbaceous herb powder by grinding dried herbaceous root with a grinder; Immersing the powder in ethanol and leaching it by stirring at a temperature of 20-30°C, filtering the leached material, and repeating the leaching and filtration 2-3 times; and concentrating the filtrate obtained by filtration under reduced pressure and freeze-drying it. The extract of Prickly pear sinensis prepared through the above process exhibits whitening activity, antioxidant activity, and anti-inflammatory activity.

Next, the method for producing a cosmetic composition according to the present invention includes preparing a fruit extract. The step of preparing the fruit extract includes washing, freeze-drying, and pulverizing tangerines, kiwis, pears, and oranges to prepare tangerine powder, kiwi powder, pear powder, and orange powder; The prepared fruit powder mixed with tangerine powder, kiwi powder, pear powder, and orange powder at a weight ratio of 3:3:2:2 was mixed with a mixed solvent of ethanol and hexane at a volume ratio of 1:1 and a weight ratio of 1:9. mixing and extracting at a temperature of 28-32°C for 22-26 hours to prepare an extract; And filtering the extract with filter paper, concentrating under reduced pressure and freeze-drying the filtered filtrate at a temperature of -48-52°C. The fruit extract is prepared through the same process as above, so that the effective ingredients can be easily extracted.

Next, the method for producing a cosmetic composition according to the present invention includes preparing a flower extract. The step of preparing the flower extract includes washing, freeze-drying, and pulverizing honeysuckle flowers, honeysuckle flowers, and peony flowers to prepare honeysuckle flower powder, honeysuckle flower powder, and peony flower powder; 20 g of the prepared mixed powder of honeysuckle flower powder, Yangha flower powder, and peony flower powder mixed at a weight ratio of 1:1:1 was mixed with 200 ml of water and stirred for 6-10 minutes at a temperature of 78-82°C. performing primary hydrothermal extraction for a period of time; After the first hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a first mixed extract; After the first hot water extraction, filter with filter paper, mix the remaining solids with water again so that the mixing ratio of solids and water is 0.1 g/ml, and do the second extraction at a temperature of 118-122°C for 10-14 hours while stirring. performing hot water extraction; After secondary hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a secondary mixed extract; After secondary hot water extraction and filtering with filter paper, the remaining solids are mixed with water again so that the mixing ratio of solids and water is 0.1 g/ml, and then mixed with [BMIm][TFSI](1-Butyl-3-methylimidazoliumbis). Adding 1-2 g of trifluoromethylsulfonyl)imide) and performing a third hot water extraction at a temperature of 98-102°C for 6-10 hours while stirring; After the third hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a third mixed extract; And mixing the first mixed extract, second mixed extract, and third mixed extract. The flower extract is prepared through the same process as above, so the active ingredients can be easily extracted.

Next, the method for producing a cosmetic composition according to the present invention includes the step of preparing grapefruit-derived exosomes. The step of preparing the grapefruit-derived exosomes includes washing the grapefruit and pretreating it at a temperature of 18-22°C and a pressure of 140-160 MPa for 3-7 minutes; Preparing grapefruit juice by squeezing pretreated grapefruit; Centrifuging the grapefruit juice to obtain a supernatant containing exosomes; Preparing a lyophilisate by lyophilizing the supernatant containing the exosomes; Forming an aqueous two-phase system using polyethylene glycol/dextran in the freeze-dried product; and obtaining a lower layer solution in which exosomes in the aqueous two-phase system are concentrated.

Next, the method for producing a cosmetic composition according to the present invention includes the step of producing a fermentation extract. The step of preparing the fermented extract includes washing corn grains and centella asiatica with boiled water and pulverizing them to prepare corn powder and centella asiatica powder; A mixture was prepared by mixing 23-27 parts by weight of corn powder, 23-27 parts by weight of Centella asiatica powder, and 58-62 parts by weight of water, and 1 part by weight of Saccharomyces cerevisiae as a yeast was added to the mixture. Fermentation at a temperature of 38-42°C for 34-38 hours; Preparing an extract by mixing the fermented mixture and butylene glycol and stirring for 4-6 days at a temperature of 28-32°C; And a step of filtering and concentrating the extract under reduced pressure and then freeze-drying it.

Next, the method for producing the cosmetic composition according to the present invention is 1-5 parts by weight of the Peach Blossom extract, 0.5-1.5 parts by weight of the fruit extract, 0.5-1.5 parts by weight of the flower extract, and 0.5-1.5 parts by weight of the grapefruit-derived exosomes. , 0.5-1.5 parts by weight of the fermented extract, 1-3 parts by weight of niacinamide, 0.5-1.5 parts by weight of arginine, 0.5-1.5 parts by weight of phenylalanine, 0.5-1.5 parts by weight of 1,2-hexanediol, pentylene glycol 0.5-1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1- 3 parts by weight, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05 It includes mixing -0.2 parts by weight, 0.05-0.2 parts by weight of polyglyceryl-6 stearate, 0.3-0.7 parts by weight of glutamic acid, and 68-72 parts by weight of carrot water.

In addition, the present invention

Provided is a health functional food composition for the prevention or improvement of melanin hyperpigmentation disease, which contains an extract of fennel root as an active ingredient and exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

In addition, the present invention

Provided is a pharmaceutical composition for the prevention or treatment of melanin hyperpigmentation disease, which contains an extract of Bunchesia chinensis as an active ingredient and exhibits skin whitening activity, antioxidant activity, and anti-inflammatory activity.

Hereinafter, the present invention will be explained in more detail by the following examples.

However, the following examples only illustrate the content of the present invention and the scope of the invention is not limited by the examples and experimental examples.

<Preparation Example 1> Preparation of fruit extract

Tangerine, kiwi, pear, and orange are washed, freeze-dried, and pulverized to prepare tangerine powder, kiwi powder, pear powder, and orange powder. The fruit powder mixed in a weight ratio of 3:3:2:2 is mixed with ethanol and hexane. An extract was prepared by mixing a mixed solvent in a volume ratio of 1:1 and a weight ratio of 1:9, and extracting the mixture at a temperature of 30°C for 24 hours. The extract was filtered through filter paper, and the filtered filtrate was concentrated under reduced pressure and freeze-dried at a temperature of -50°C to prepare a fruit extract.

<Preparation Example 2> Preparation of flower extract

Honeysuckle flowers, Yangha flowers, and peony flowers were washed, freeze-dried, and pulverized to prepare honeysuckle flower powder, Yangha flower powder, and peony flower powder, and a mixed powder was prepared by mixing them at a weight ratio of 1:1:1. 20 g of the mixed powder was mixed with 200 ml of water, and primary hot water extraction was performed at a temperature of 80° C. for 8 hours while stirring. Then, the mixture was filtered through filter paper and the filtrate was freeze-dried to prepare a primary mixed extract. .

After the first hot water extraction, filter through filter paper and mix the remaining solids with water again so that the mixing ratio of solids and water is 0.1 g/ml, and perform the second hot water extraction at a temperature of 120°C for 12 hours while stirring. Afterwards, it was filtered through filter paper and the filtrate was freeze-dried to prepare a secondary mixed extract.

After secondary hot water extraction and filtering with filter paper, the remaining solids are mixed with water again so that the mixing ratio of solids and water is 0.1 g/ml, and then mixed with [BMIm][TFSI](1-Butyl-3-methylimidazoliumbis). After adding 1 g of trifluoromethylsulfonyl)imide), a third hot water extraction was performed at a temperature of 100°C for 8 hours while stirring, filtered through filter paper, and the filtrate was freeze-dried to prepare a third mixed extract.

A flower extract was prepared by mixing the first mixed extract, second mixed extract, and third mixed extract.

<Preparation Example 3> Preparation of grapefruit-derived exosomes

Grapefruit was washed and pretreated at a temperature of 20°C and a pressure of 150 MPa for 5 minutes, and the pretreated grapefruit was squeezed to prepare grapefruit juice. The grapefruit juice was centrifuged to obtain a supernatant containing exosomes, and the supernatant containing the exosomes was lyophilized to prepare a freeze-dried product. Polyethylene glycol/dextran was used in the freeze-dried material to form an aqueous two-phase system, and a lower layer containing concentrated exosomes in the aqueous two-phase system was obtained to prepare grapefruit-derived exosomes.

<Preparation Example 4> Fermentation extract production

Corn grains and Centella asiatica were washed with boiled water and pulverized to prepare corn powder and Centella asiatica powder. A mixture was prepared by mixing 25 parts by weight of corn powder, 25 parts by weight of Centella asiatica powder, and 60 parts by weight of water, and the mixture was infused with yeast. 1 part by weight of Saccharomyces cerevisiae was added and fermented at a temperature of 40°C for 36 hours. An extract was prepared by adding 60 parts by weight of butylene glycol to 100 parts by weight of the fermented mixture and stirring for 5 days at a temperature of 30°C. The extract was filtered, concentrated under reduced pressure, and then freeze-dried to prepare a fermented extract.

<Example 1> Manufacture of Bunhaengcho raw extract (Bunhaengcho extract Ⅰ)

Beonhaengcho, a halophyte native to Jeju, was provided by Dongan Co., Ltd. and used in the experiment.

Prepare the raw Herbaceous herb by cutting it with scissors, weigh a certain amount of the prepared Herbaceous herb raw material, add 10 times 70% (v/v) ethanol, stir and leach at a temperature of 25°C for 2.5 days, and the leached leachate. Filtration was performed, and this leaching and filtration process was repeated three times, and the obtained filtrate was concentrated under reduced pressure and freeze-dried to prepare an extract of Bunhaengcho.

Figure 1 shows the extract manufacturing process in photographs.

<Example 2> Preparation of dried extract of Bunhaengcho extract (Bunhaengcho extract Ⅱ)

Beonhaengcho, a halophyte native to Jeju, was provided by Dongan Co., Ltd. and used in the experiment.

The raw material of Bunhaengcho was dried at a temperature of 50°C for 3 hours, and the dried Bunhaengcho was pulverized with a grinder to prepare Bunhaengcho powder. After weighing a certain amount of the above powder, 70% (v/v) ethanol was added 10 times, leached by stirring at a temperature of 25°C for 2.5 days, and the leached leachate was filtered. This leaching and filtration process was carried out in 3 steps. This was repeated several times, and the obtained filtrate was concentrated under reduced pressure and freeze-dried to prepare an extract of Bunhaengcho.

Figure 1 shows the extract manufacturing process in photographs.

<Comparative Example 1> Preparation of Namunjae extract

Namunjae, a halophyte native to Jeju, was provided by Dongan Co., Ltd. and used in the experiment.

The raw wood material was dried at a temperature of 50°C for 3 hours, and the dried wood material was pulverized with a grinder to prepare bare wood powder. After weighing a certain amount of the above-mentioned bare material powder, 10 times 70% (v/v) ethanol was added, leached by stirring at a temperature of 25°C for 2.5 days, and the leached leachate was filtered. This leaching and filtration process was carried out in 3 steps. This was repeated several times, and the obtained filtrate was concentrated under reduced pressure and freeze-dried to prepare a Namunjae extract.

Figure 1 shows the extract manufacturing process in photographs.

<Example 3> Preparation of whitening cosmetic composition-1

3 parts by weight of the P. lily extract prepared in Example 2, 2 parts by weight of niacinamide, 1 part by weight of arginine, 1 part by weight of phenylalanine, 1 part by weight of 1,2-hexanediol, 1 part by weight of pentylene glycol, glycerin 1 part by weight of lyl caprylate, 1 part by weight of ascorbyl glucoside, 5 parts by weight of squalane, 2 parts by weight of dextrin, 2 parts by weight of linalool, 2 parts by weight of limonene, 2 parts by weight of xanthan gum, 1 part by weight of adenosine, 0.1 part by weight of stearyl alcohol Part by weight, 0.1 part by weight of sodium hyaluronate, 0.1 part by weight of valine, 0.1 part by weight of isoquercitrin, 0.1 part by weight of polyglyceryl-6 stearate, 0.5 part by weight of glutamic acid and 74 parts by weight of carrot water are mixed. A cosmetic composition for whitening was prepared.

<Example 4> Preparation of whitening cosmetic composition-2

3 parts by weight of the Bunhaengcho extract prepared in Example 2, 1 part by weight of the fruit extract prepared in Preparation Example 1, 1 part by weight of the flower extract prepared in Preparation Example 2, grapefruit-derived exosomes prepared in Preparation Example 3 1 part by weight, 1 part by weight of fermented extract prepared in Preparation Example 4, 2 parts by weight of niacinamide, 1 part by weight of arginine, 1 part by weight of phenylalanine, 1 part by weight of 1,2-hexanediol, 1 part by weight of pentylene glycol Parts by weight, 1 part by weight of glyceryl caprylate, 1 part by weight of ascorbyl glucoside, 5 parts by weight of squalane, 2 parts by weight of dextrin, 2 parts by weight of linalool, 2 parts by weight of limonene, 2 parts by weight of xanthan gum, 1 part by weight of adenosine, 0.1 part by weight of stearyl alcohol, 0.1 part by weight of sodium hyaluronate, 0.1 part by weight of valine, 0.1 part by weight of isoquercitrin, 0.1 part by weight of polyglyceryl-6 stearate, 0.5 part by weight of glutamic acid, and 70 parts by weight of carrot water. A whitening cosmetic composition was prepared by mixing parts by weight.

<Experimental Example 1> Total polyphenol content analysis

The total polyphenol content of the extracts prepared in Example 1, Example 2, and Comparative Example 1 was confirmed. Total polyphenol content was measured by applying the Folin-Denis method. Equal amounts of the extract diluted by concentration and Folin reagent diluted twice were mixed and left for 3 minutes. Then, an equal amount of 10% Na2CO ₃ was added and reacted for 1 hour. Then, the absorbance was measured at 700 nm using a Microplate Spectrophotometer, and the standard curve was prepared. Total polyphenol content was determined. At this time, a standard curve was obtained using gallic acid, and the results are shown in Figure 2.

Polyphenol compounds, which are widely distributed in the plant world, are well known not only as natural pigments but also as natural antioxidants that remove oxygen radicals generated in the living body. Polyphenol compounds are aromatic compounds with two or more phenolic hydroxyl groups in one molecule, and there are more than 1,000 different types, including not only phenolic acids but also cinamic acid derivatives, anthocyanins, flavonoids, lignans, resveratrol, and tannins. Polyphenol compounds are important phytochemicals that not only have antioxidant, but also anti-cancer, anti-diabetic, antibacterial, anti-allergic, anti-hypertensive, and anti-aging activities. It's in the spotlight.

The polyphenol contents of the Bunhaengcho extract prepared in Example 1 (Bunhaengcho extract I), the Bunhaengcho extract prepared in Example 2 (Bunhaengcho extract II), and the Namunjae extract prepared in Comparative Example 1 were 15.32, 20.24, and 17.01, respectively. It was confirmed that the extract of Bunhaengcho of Example 2 contained the highest content of polyphenol in mg GAE/g.

<Experimental Example 2> 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging activity

The antioxidant activity of the extracts prepared in Examples 1, 2, and Comparative Example 1 was confirmed. Measurement of antioxidant power using ABTS radical was performed using the ABTS+·cation decolorization assay method. Mix equal amounts of 7mM ABTS and 2.45mM potassium persulfate to the final concentration, react at room temperature in the dark for 24 hours to form ABTS+·, and then add phosphate buffered saline (PBS, pH7) to an absorbance value of 0.70 (±0.02) at 732 nm. It was diluted to .4). 180 μl of sample was mixed with 20 μl of diluted ABTS solution and left for 1 minute to measure absorbance. The results are shown in Table 1 and Figure 3 below.

division Concentration (㎍/mL) Scavenging effect (%) Example 1
(Bunhaengcho ExtractⅠ) 31.25 15.83±0.33 62.5 35.57±0.41 125 42.87±0.88 250 72.37±0.56 500 90.67±0.31 Example 2
(Bunhaengcho extract Ⅱ) 31.25 13.68±2.04 62.5 27.93±3.90 125 48.57±9.36 250 84.31±1.80 500 96.32±2.12 Comparative Example 1
(Namunjae Extract) 31.25 13.21±7.66 62.5 27.98±10.48 125 45.05±0.59 250 78.52±0.99 500 92.72±0.09 Vit.C One 6.07±1.32 2.5 11.56±0.87 5 34.97±1.73 7.5 95.09±0.50

The ABTS radical scavenging activity of the Bunhaengcho extract prepared in Example 1 (Bunhaengcho extract I), the Bunhaengcho extract prepared in Example 2 (Bunhaengcho extract II), and the Namunjae extract prepared in Comparative Example 1 were tested with ascorbic acid (Vit. As shown in Figure 3 as a result of measurement by concentration compared to C), Example 2 showed excellent scavenging activity of 84.31% at a concentration of 250 ㎍/mL. The ABTS radical RC ₅₀ values of Example 1, Example 2, and Comparative Example 1 were confirmed to be 155.32 ㎍/mL, 138.22 ㎍/mL, and 147.77 ㎍/mL, respectively, showing that the extract of P. syringa vulgaris prepared in Example 2 had the highest radical scavenging activity. It was confirmed to be excellent. In particular, it showed an improved DPPH radical inhibitory activity of about 6.47% compared to the Namunjae extract prepared in Comparative Example 1.

<Experimental Example 3> DPPH radical scavenging activity

The antioxidant activity of the extracts prepared in Examples 1, 2, and Comparative Example 1 was confirmed. Free radical scavenging activity was measured using the principle of measuring the reducing power of DPPH, a stable radical. The initial value of 160 ㎕ of each concentration of extract diluted in 99% methanol was measured at 517 nm, then mixed with 40 ㎕ of 0.15 mM DPPH solution dissolved in methanol and left in the dark for 30 minutes, and then the absorbance was measured at 517 nm. DPPH radical scavenging activity was calculated according to the formula below, methanol was used as a control, and the results are shown in Table 2 and Figure 4.

<expression>

DPPH radical scavenging activity (%) = [100-(S/C×100)]

S: Absorbance after sample group reaction - Absorbance before sample group reaction

C: Absorbance after control reaction - Absorbance before control reaction

division Concentration (㎍/mL) Scavenging effect (%) Example 1
(Bunhaengcho ExtractⅠ) 31.25 2.12±3.61 62.5 5.03±1.50 125 11.68±0.50 250 48.96±0.50 500 80.17±1.32 Example 2
(Bunhaengcho extract Ⅱ) 31.25 6.36±0.87 62.5 14.45±4.28 125 20.32±3.13 250 69.72±1.80 500 99.25±1.50 Comparative Example 1
(Namunjae Extract) 31.25 2.72±2.18 62.5 7.75±4.78 125 18.61±4.78 250 52.57±2.50 500 92.50±0.87 Vit.C One 13.42±3.97 2.5 31.83±1.25 5 51.90±9.30 7.5 93.86±4.62

DPPH radical of the Bunhaengcho extract prepared in Example 1 (Beonhaengcho extract I), the Bunhaengcho extract prepared in Example 2 (Beonhaengcho extract II), and the Namunjae extract prepared in Comparative Example 1 and ascorbic acid (Vit.C) The results of measuring and comparing scavenging activity at different concentrations are shown in Figure 4. Ascorbic acid (Vit.C) used as a positive control showed antioxidant activity of more than 90% at 7.5 ㎍/mL, and in Example 2, it had excellent scavenging activity of 69.72% at a concentration of 250 ㎍/mL. The RC ₅₀ values of Example 1, Example 2, and Comparative Example 1 were confirmed to be 267.37 ㎍/mL, 192.64 ㎍/mL, and 237.52 ㎍/mL, respectively, showing that the radical scavenging activity was the most excellent in the extract of Hungrye sinensis prepared in Example 2. Confirmed. In particular, it showed an improved DPPH radical inhibitory activity of about 18% compared to the Namunjae extract prepared in Comparative Example 1.

<Experimental Example 4> Cytotoxicity and anti-inflammatory activity analysis

The cytotoxic and anti-inflammatory activities of the extracts prepared in Example 1, Example 2, and Comparative Example 1 were confirmed.

- Cell line culture

RAW 264.7 (mouse macrophage cell line) cells used in the experiment were purchased from the Korean Cell Line Bank. RAW264.7 cells were subcultured once every 2-3 days in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 100 ㎍/mL penicillin in an incubator at 37°C with 95% oxygen and 5% CO ₂ . I did it.

- Measurement of cell viability

To confirm the cell viability of Example 1 (Bunhaengcho extract Ⅰ), Example 2 (Bunhaengcho extract Ⅱ), and Comparative Example 1 (Namunjae extract) in RAW 264.7 cells, 3-(4,5-Dimethylthiazol-2-yl)- Cell viability was measured with 2,5-Diphenyltetrazolium Bromide (MTT). Cells (5 × 10 ⁴ /well) were dispensed at 100 ㎕ each into a 96-well plate and cultured in a CO ₂ incubator for 24 hours. Then, replaced with seru-free medium, samples were treated at each concentration and 5 mg/mL after 24 hours. 10 ㎕ of MTT was dispensed and reacted for 4 hours to reduce MTT. Afterwards, the supernatant was removed, and dimethyl sulfoxide (DMSO) was dispensed in 100 ㎕ portions to dissolve the formazone cell crystals, and the absorbance was measured at 540 nm using a Microplate Spectrophotometer. Cell viability was expressed as a percentage (%) compared to the control group.

Lipopolysaccharide (LPS), a cell wall material of Gram-negative bacteria, has been reported to stimulate immune cells and increase NO production. The cell survival rate when RAW 264.7 cells induced by oxidative stress with LPS were treated with the extracts of Examples 1, 2, and Comparative Example 1 at different concentrations and cultured for a certain period of time is shown in Figure 5. It was confirmed that there was no cytotoxicity due to LPS stimulation. In the case of the extracts of Example 1, Example 2, and Comparative Example 1, cell viability was more than 90% at all concentrations of 31.25, 62.5, 125, 250, and 500 μg/mL, confirming that there was no toxicity to cells.

- Inhibition of NO production

NO content was measured by the method of Green et al. RAW 264.7 cells were distributed in a 96 well plate at a concentration of 1×10 ⁵ cells/mL using DMEM medium, and then simultaneously treated with new medium containing the test sample and LPS (100 ng/mL, Sigma Co.) for 24 days. cultured for some time. 100 ㎕ of cell culture supernatant and 100 ㎕ of Griess reagent (1% sulfanilamide, 0.1% naphthylethylendiaminein, 2.5% phosphoric acid) were mixed and reacted for 10 minutes in a 96 well plate. Absorbance was measured at 540 nm using an ELISA autoreader, and sodium nitrate ( The content of NO was calculated by creating a standard curve using Sigma Co.).

NO produced by LPS stimulation mediates important physiological functions such as vascular homeostasis and apoptosis induction under normal physiological conditions, but large amounts of NO kill normal cells and induce inflammation, causing acute or chronic inflammatory diseases. It works. Therefore, effective NO secretion control is known as a treatment method for acute or chronic inflammatory diseases, and research on this is actively underway.

Accordingly, as a result of examining whether the extracts of Example 1 (Bunhaengcho extract I), Example 2 (Bunhaengcho extract II), and Comparative Example 1 (Namunjae extract) could inhibit the production of NO, the results were shown in Figure 6. When measuring the toxicity of the extracts to cells using the MTT assay before measuring the effect of inhibiting NO production, all three extracts showed a cell viability of more than 100% at a concentration of 500 ㎍/mL, so the extract concentrations were 62.5 and 62.5, respectively. Cells were treated at concentrations of 125, 250, and 500 μg/mL. As a result, in the group treated only with LPS, NO was produced at about 31.24 μM, which is about 5 times more NO than the untreated group, and Example 1 (Bunhaengcho extract I), Example 2 (Bunhaengcho extract II), and Comparative Example When treated with the extract of 1 (Namunjae extract), significant NO scavenging activity was shown starting at a concentration of 250 ㎍/mL.

In particular, the extract of Bunhaengcho of Example 2 showed significant scavenging activity at a concentration of 125 ㎍/mL, and when compared with the Namunjae extract of Comparative Example 1 at the same concentration of 250 ㎍/mL, the scavenging activity increased by about 32%. was confirmed.

<Experimental Example 5> Whitening activity analysis-1

The whitening activity of the extract prepared in Example 2 was confirmed.

- Cell line culture

The B16F10 melanoma cell line used in the experiment was purchased from ATCC (Virginia, USA) and cultured in DMEM medium containing 10% heat-inactivated FBS, 1% penicillin/streptomycin, and 1.5 g/L sodium bicarbonate at 37°C and 5% CO ^2. did.

- Experiment to check melanin production amount

B16F10 cells were distributed in a 6-well plate at a density of 5×10 ⁴ cells/well and cultured for 1 day. Then, the cells were cultured for 3 days in a medium containing 2% FBS, treated with different concentrations of extract. To induce melanin production, 200 nM α-MSH was co-treated, and 200 μM arbutin was used as a positive control. After completion of the culture, the cells were recovered and lysed by treating them with 200 ㎕ of 1 M NaOH. The absorbance at 490 nm of the lysate was measured using a spectrophotometer.

To confirm the melanin synthesis inhibitory effect of the extract prepared in Example 2, the melanin production rate was compared after treating B16F10 cells at concentrations of 100, 200, and 400 μg/mL, and the results are shown in Figure 7.

As shown in Figure 7, concentration-dependent inhibition of melanin production by the extract of Bunhaengcho extract prepared in Example 2 was observed.

<Experimental Example 6> Whitening activity analysis-2

The whitening activity of the cosmetic composition prepared in Example 3-4 was confirmed.

Using the above cosmetic composition, 36 healthy women aged 30-40 were randomly divided into two groups. After washing their face twice every morning and evening with the cosmetic composition of Example 1-2, an appropriate amount was applied to the left side of the face and the right side of the face. It was applied continuously for months. The whitening effect was evaluated by observing the degree of facial improvement one month after the start of the experiment and after the end of the experiment, and measuring the number of test subjects who showed the effect. The experimental results are shown in Table 3 below.

When to observe division available
(number of people) slightly valid
(number of people) invalidity
(number of people) effective rate
(%) 1 month later Example 3 28 6 2 94.4 Example 4 30 6 0 100.0 After the experiment is over Example 3 30 5 One 97.2 Example 4 32 4 0 100.0

As shown in Table 3, it was confirmed that the cosmetic composition according to the present invention had an excellent whitening effect.

Claims (7)

1-5 parts by weight of Prickly pear extract, 0.5-1.5 parts by weight of fruit extract mixed with tangerine extract, kiwi extract, pear extract, and orange extract in a weight ratio of 3:3:2:2, honeysuckle extract, honeysuckle flower extract, and 0.5-1.5 parts by weight of flower extract mixed with peony flower extract at a weight ratio of 1:1:1, 0.5-1.5 parts by weight of grapefruit-derived exosomes, fermented corn extract, and fermented centella asiatica extract mixed at a weight ratio of 1:1. Fermentation extract 0.5-1.5 parts by weight, niacinamide 1-3 parts by weight, arginine 0.5-1.5 parts by weight, phenylalanine 0.5-1.5 parts by weight, 1,2-hexanediol 0.5-1.5 parts by weight, pentylene glycol 0.5- 1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1-3 parts by weight parts, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05-0.2 A cosmetic composition comprising 0.05-0.2 parts by weight of polyglyceryl-6stearate, 0.3-0.7 parts by weight of glutamic acid, and 68-72 parts by weight of carrot water. Drying the raw material of Tetragonia tetragonioides at a temperature of 40-60°C; Preparing dried herbaceous herb powder by grinding dried herbaceous root with a grinder; Immersing the powder in ethanol and leaching it by stirring at a temperature of 20-30°C, filtering the leached material, and repeating the leaching and filtration 2-3 times; and concentrating the filtrate obtained by filtration under reduced pressure and freeze-drying; preparing an extract of Bunhaengcho, including;
Washing, freeze-drying, and pulverizing tangerines, kiwis, pears, and oranges to prepare tangerine powder, kiwi powder, pear powder, and orange powder; The prepared fruit powder mixed with tangerine powder, kiwi powder, pear powder, and orange powder at a weight ratio of 3:3:2:2 was mixed with a mixed solvent of ethanol and hexane at a volume ratio of 1:1 and a weight ratio of 1:9. mixing and extracting at a temperature of 28-32°C for 22-26 hours to prepare an extract; And filtering the extract through filter paper, concentrating and freeze-drying the filtered filtrate under reduced pressure at a temperature of -48-52°C; Preparing a fruit extract comprising a;
Preparing honeysuckle flower powder, Yangha flower powder, and peony flower powder by washing, freeze-drying, and pulverizing honeysuckle flowers, Yangha flowers, and peony flowers; 20 g of the prepared mixed powder of honeysuckle flower powder, Yangha flower powder, and peony flower powder mixed at a weight ratio of 1:1:1 was mixed with 200 ml of water and stirred for 6-10 minutes at a temperature of 78-82°C. performing primary hydrothermal extraction for a period of time; After the first hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a first mixed extract; After the first hot water extraction, filter with filter paper, mix the remaining solids with water again so that the mixing ratio of solids and water is 0.1 g/ml, and do the second extraction at a temperature of 118-122°C for 10-14 hours while stirring. performing hot water extraction; After secondary hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a secondary mixed extract; After secondary hot water extraction and filtering with filter paper, the remaining solids are mixed with water again so that the mixing ratio of solids and water is 0.1 g/ml, and then mixed with [BMIm][TFSI](1-Butyl-3-methylimidazoliumbis). Adding 1-2 g of trifluoromethylsulfonyl)imide) and performing a third hot water extraction at a temperature of 98-102°C for 6-10 hours while stirring; After the third hot water extraction, filtering through filter paper and then freeze-drying the filtrate to prepare a third mixed extract; And mixing the first mixed extract, second mixed extract, and third mixed extract; Preparing a flower extract comprising a;
Washing the grapefruit and pretreating it at a temperature of 18-22°C and a pressure of 140-160 MPa for 3-7 minutes; Preparing grapefruit juice by squeezing pretreated grapefruit; Centrifuging the grapefruit juice to obtain a supernatant containing exosomes; Preparing a lyophilisate by lyophilizing the supernatant containing the exosomes; Forming an aqueous two-phase system using polyethylene glycol/dextran in the freeze-dried product; And obtaining a lower layer liquid in which exosomes are concentrated in the aqueous two-phase system; Preparing grapefruit-derived exosomes comprising;
Preparing corn powder and Centella asiatica powder by washing corn grains and Centella asiatica with boiled water and pulverizing them; A mixture was prepared by mixing 23-27 parts by weight of corn powder, 23-27 parts by weight of Centella asiatica powder, and 58-62 parts by weight of water, and 1 part by weight of Saccharomyces cerevisiae as a yeast was added to the mixture. Fermentation at a temperature of 38-42°C for 34-38 hours; Preparing an extract by mixing the fermented mixture and butylene glycol and stirring for 4-6 days at a temperature of 28-32°C; and filtering and concentrating the extract under reduced pressure and then freeze-drying the extract. Preparing a fermentation extract including; and
1-5 parts by weight of the Bunhaengcho extract, 0.5-1.5 parts by weight of the fruit extract, 0.5-1.5 parts by weight of the flower extract, 0.5-1.5 parts by weight of the grapefruit-derived exosomes, 0.5-1.5 parts by weight of the fermented extract, niacinamide 1 -3 parts by weight, arginine 0.5-1.5 parts by weight, phenylalanine 0.5-1.5 parts by weight, 1,2-hexanediol 0.5-1.5 parts by weight, pentylene glycol 0.5-1.5 parts by weight, glyceryl caprylate 0.5-1.5 parts by weight Parts by weight, ascorbyl glucoside 0.5-1.5 parts by weight, squalane 3-7 parts by weight, dextrin 1-3 parts by weight, linalool 1-3 parts by weight, limonene 1-3 parts by weight, xanthan gum 1-3 parts by weight, adenosine 0.5-1.5 parts by weight, stearyl alcohol 0.05-0.2 parts by weight, sodium hyaluronate 0.05-0.2 parts by weight, valine 0.05-0.2 parts by weight, isoquercitrin 0.05-0.2 parts by weight, polyglyceryl-6 stearate 0.05 - A method of producing a cosmetic composition comprising mixing 0.2 parts by weight, 0.3-0.7 parts by weight of glutamic acid, and 68-72 parts by weight of carrot water. delete delete delete delete delete